Journal: Nature chemical biology

Article Title: Bidirectional Modulation of HIF-2 Activity through Chemical Ligands

doi: 10.1038/s41589-019-0234-5

Figure Lengend Snippet: a , Chemical structure of M1002. b , Clear agonistic effects on the expression of HIF-2 target genes in 786-O cells by M1002 (10 μM) as compared with the antagonist PT2385 (1 μM). c , TR-FRET-based protein binding assays testing the effects of M1002 on the associations between ARNT and its partners HIF-2α, HIF-1α and NPAS3, respectively. d , Comparison of M1002’s agonistic effects at 10 μM on HIF-2 target genes in HEK293T cells transfected with wild-type HIF-2α or Y281A mutant. Error bars, mean ± s.d.; n=3 (distinct replicates for cell cultures). e , Co-IP results showing the effects of HIF-2α ligands (PT2385, T1001, M1001 and M1002 at various concentrations) on the association between full-length HIF-2α and VHL proteins. This experiment was independently repeated twice with similar results.

Article Snippet: For the co-treatment of PHD inhibitors and HIF-2α agonist, Hep3B cells were incubated with 10 μM of molidustat (Cayman Chemical 15297) or roxadustat (Cayman Chemical 15294) for 8 h before 5 μM of M1002 (Molport 001–006-026, >95% purity) was added into the medium (final DMSO concentration all kept at 0.1%).

Techniques: Expressing, Protein Binding, Comparison, Transfection, Mutagenesis, Co-Immunoprecipitation Assay